Source 1primary paper2020Journal of the American Society for Mass Spectrometry
Toolkit/electron-transfer/higher-energy collision dissociation
electron-transfer/higher-energy collision dissociation
Also known as: EThcD
Taxonomy: Technique Branch / Method. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
Electron-transfer/higher-energy collision dissociation (EThcD) is a top-down mass spectrometry fragmentation method used in a combined workflow with 213 nm ultraviolet photodissociation (UVPD) to characterize covalent insulin dimers. In the cited study, this workflow identified cross-link chemical composition and, with MS3 analysis of informative MS2 fragments, enabled residue-level localization of interchain cross-link sites.
Usefulness & Problems
Why this is useful
EThcD is useful for structural characterization of covalent protein dimers when cross-link identity and linkage position must both be resolved. In the cited insulin study, combining EThcD with 213 nm UVPD improved interpretation of cross-linked species by supporting both cross-link composition assignment and site localization.
Source:
The combined utilization of EThcD and 213 nm ultraviolet photodissociation (UVPD) facilitated identification of the chemical composition of the cross-links.
Source:
Identification of cross-link sites between chains at residue level was achievable for both dimers with MS3 analysis of MS2 fragments cleaved at the cross-link or additionally the interchain disulfide bonds.
Source:
we here demonstrate on two case studies of covalent insulin dimers, induced by Fe2+ incubation or ultraviolet (UV) light stress, that de novo characterization in top-down mass spectrometry (MS) workflows can identify cross-link types and sites
Problem solved
This method addresses the analytical problem of determining both the chemical nature and residue-level positions of covalent cross-links in insulin dimers formed under Fe2+ incubation or UV light stress. It is particularly relevant when a single MS2 fragmentation mode is insufficient for complete cross-link site identification, as observed for the UV light-induced dimer.
Problem links
Need precise spatiotemporal control with light input
DerivedElectron-transfer/higher-energy collision dissociation (EThcD) is a top-down mass spectrometry fragmentation method used in combination with 213 nm ultraviolet photodissociation (UVPD) to characterize covalent insulin dimers. In the cited study, combined EThcD and UVPD enabled identification of cross-link chemical composition, and MS3 analysis of MS2 fragments supported residue-level localization of interchain cross-link sites.
Taxonomy & Function
Primary hierarchy
Technique Branch
Method: A concrete measurement method used to characterize an engineered system.
Mechanisms
electron-transfer dissociationelectron-transfer dissociationHeterodimerizationhigher-energy collision-induced dissociationhigher-energy collision-induced dissociationPhotocleavagePhotocleavageultraviolet photodissociationultraviolet photodissociationTechniques
Functional AssayTarget processes
No target processes tagged yet.
Input: Light
Implementation Constraints
cofactor dependency: cofactor requirement unknownencoding mode: genetically encodedimplementation constraint: context specific validationimplementation constraint: multi component delivery burdenimplementation constraint: spectral hardware requirementoperating role: sensorswitch architecture: cleavageswitch architecture: multi componentswitch architecture: recruitment
The validated implementation used a top-down, multistage tandem mass spectrometry workflow combining EThcD with 213 nm UVPD. Residue-level site assignment relied on MS3 analysis of selected MS2 fragments, including fragments produced by cleavage at the cross-link and, in some cases, at interchain disulfide bonds.
The evidence is limited to a single study on covalent insulin dimers, so broader generalizability to other proteins or cross-link classes is not established. MS2 alone was not sufficient for cross-link site identification in the UV light-induced dimer, indicating that multistage analysis may be required for some analytes.
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Supporting Sources
Ranked Claims
UVPD at MS2 provided cross-link site identification for the Fe2+-induced insulin dimer without MS3, but MS2 was not sufficient for cross-link site identification in the UV light-induced dimer.
UVPD provided identification of cross-link sites in the Fe2+-induced dimer without MS3, while cross-link site identification with MS2 was not possible for the UV light-induced dimer.
UVPD at MS2 provided cross-link site identification for the Fe2+-induced insulin dimer without MS3, but MS2 was not sufficient for cross-link site identification in the UV light-induced dimer.
UVPD provided identification of cross-link sites in the Fe2+-induced dimer without MS3, while cross-link site identification with MS2 was not possible for the UV light-induced dimer.
UVPD at MS2 provided cross-link site identification for the Fe2+-induced insulin dimer without MS3, but MS2 was not sufficient for cross-link site identification in the UV light-induced dimer.
UVPD provided identification of cross-link sites in the Fe2+-induced dimer without MS3, while cross-link site identification with MS2 was not possible for the UV light-induced dimer.
UVPD at MS2 provided cross-link site identification for the Fe2+-induced insulin dimer without MS3, but MS2 was not sufficient for cross-link site identification in the UV light-induced dimer.
UVPD provided identification of cross-link sites in the Fe2+-induced dimer without MS3, while cross-link site identification with MS2 was not possible for the UV light-induced dimer.
UVPD at MS2 provided cross-link site identification for the Fe2+-induced insulin dimer without MS3, but MS2 was not sufficient for cross-link site identification in the UV light-induced dimer.
UVPD provided identification of cross-link sites in the Fe2+-induced dimer without MS3, while cross-link site identification with MS2 was not possible for the UV light-induced dimer.
UVPD at MS2 provided cross-link site identification for the Fe2+-induced insulin dimer without MS3, but MS2 was not sufficient for cross-link site identification in the UV light-induced dimer.
UVPD provided identification of cross-link sites in the Fe2+-induced dimer without MS3, while cross-link site identification with MS2 was not possible for the UV light-induced dimer.
UVPD at MS2 provided cross-link site identification for the Fe2+-induced insulin dimer without MS3, but MS2 was not sufficient for cross-link site identification in the UV light-induced dimer.
UVPD provided identification of cross-link sites in the Fe2+-induced dimer without MS3, while cross-link site identification with MS2 was not possible for the UV light-induced dimer.
UVPD at MS2 provided cross-link site identification for the Fe2+-induced insulin dimer without MS3, but MS2 was not sufficient for cross-link site identification in the UV light-induced dimer.
UVPD provided identification of cross-link sites in the Fe2+-induced dimer without MS3, while cross-link site identification with MS2 was not possible for the UV light-induced dimer.
UVPD at MS2 provided cross-link site identification for the Fe2+-induced insulin dimer without MS3, but MS2 was not sufficient for cross-link site identification in the UV light-induced dimer.
UVPD provided identification of cross-link sites in the Fe2+-induced dimer without MS3, while cross-link site identification with MS2 was not possible for the UV light-induced dimer.
UVPD at MS2 provided cross-link site identification for the Fe2+-induced insulin dimer without MS3, but MS2 was not sufficient for cross-link site identification in the UV light-induced dimer.
UVPD provided identification of cross-link sites in the Fe2+-induced dimer without MS3, while cross-link site identification with MS2 was not possible for the UV light-induced dimer.
The UV chromophoric side chain of Phe1 was implicated in the Fe2+-induced dimer cross-link, explaining why UVPD-MS2 effectively fragmented the cross-link and nearby backbone bonds.
The UV chromophoric side chain of Phe1 was indicated in the cross-link, explaining why UVPD-MS2 was effective in fragmenting the cross-link and nearby backbone bonds.
The UV chromophoric side chain of Phe1 was implicated in the Fe2+-induced dimer cross-link, explaining why UVPD-MS2 effectively fragmented the cross-link and nearby backbone bonds.
The UV chromophoric side chain of Phe1 was indicated in the cross-link, explaining why UVPD-MS2 was effective in fragmenting the cross-link and nearby backbone bonds.
The UV chromophoric side chain of Phe1 was implicated in the Fe2+-induced dimer cross-link, explaining why UVPD-MS2 effectively fragmented the cross-link and nearby backbone bonds.
The UV chromophoric side chain of Phe1 was indicated in the cross-link, explaining why UVPD-MS2 was effective in fragmenting the cross-link and nearby backbone bonds.
The UV chromophoric side chain of Phe1 was implicated in the Fe2+-induced dimer cross-link, explaining why UVPD-MS2 effectively fragmented the cross-link and nearby backbone bonds.
The UV chromophoric side chain of Phe1 was indicated in the cross-link, explaining why UVPD-MS2 was effective in fragmenting the cross-link and nearby backbone bonds.
The UV chromophoric side chain of Phe1 was implicated in the Fe2+-induced dimer cross-link, explaining why UVPD-MS2 effectively fragmented the cross-link and nearby backbone bonds.
The UV chromophoric side chain of Phe1 was indicated in the cross-link, explaining why UVPD-MS2 was effective in fragmenting the cross-link and nearby backbone bonds.
The UV chromophoric side chain of Phe1 was implicated in the Fe2+-induced dimer cross-link, explaining why UVPD-MS2 effectively fragmented the cross-link and nearby backbone bonds.
The UV chromophoric side chain of Phe1 was indicated in the cross-link, explaining why UVPD-MS2 was effective in fragmenting the cross-link and nearby backbone bonds.
The UV chromophoric side chain of Phe1 was implicated in the Fe2+-induced dimer cross-link, explaining why UVPD-MS2 effectively fragmented the cross-link and nearby backbone bonds.
The UV chromophoric side chain of Phe1 was indicated in the cross-link, explaining why UVPD-MS2 was effective in fragmenting the cross-link and nearby backbone bonds.
The UV chromophoric side chain of Phe1 was implicated in the Fe2+-induced dimer cross-link, explaining why UVPD-MS2 effectively fragmented the cross-link and nearby backbone bonds.
The UV chromophoric side chain of Phe1 was indicated in the cross-link, explaining why UVPD-MS2 was effective in fragmenting the cross-link and nearby backbone bonds.
The UV chromophoric side chain of Phe1 was implicated in the Fe2+-induced dimer cross-link, explaining why UVPD-MS2 effectively fragmented the cross-link and nearby backbone bonds.
The UV chromophoric side chain of Phe1 was indicated in the cross-link, explaining why UVPD-MS2 was effective in fragmenting the cross-link and nearby backbone bonds.
The UV chromophoric side chain of Phe1 was implicated in the Fe2+-induced dimer cross-link, explaining why UVPD-MS2 effectively fragmented the cross-link and nearby backbone bonds.
The UV chromophoric side chain of Phe1 was indicated in the cross-link, explaining why UVPD-MS2 was effective in fragmenting the cross-link and nearby backbone bonds.
Combined EThcD and 213 nm UVPD facilitated identification of the chemical composition of cross-links in insulin dimers.
The combined utilization of EThcD and 213 nm ultraviolet photodissociation (UVPD) facilitated identification of the chemical composition of the cross-links.
Combined EThcD and 213 nm UVPD facilitated identification of the chemical composition of cross-links in insulin dimers.
The combined utilization of EThcD and 213 nm ultraviolet photodissociation (UVPD) facilitated identification of the chemical composition of the cross-links.
Combined EThcD and 213 nm UVPD facilitated identification of the chemical composition of cross-links in insulin dimers.
The combined utilization of EThcD and 213 nm ultraviolet photodissociation (UVPD) facilitated identification of the chemical composition of the cross-links.
Combined EThcD and 213 nm UVPD facilitated identification of the chemical composition of cross-links in insulin dimers.
The combined utilization of EThcD and 213 nm ultraviolet photodissociation (UVPD) facilitated identification of the chemical composition of the cross-links.
Combined EThcD and 213 nm UVPD facilitated identification of the chemical composition of cross-links in insulin dimers.
The combined utilization of EThcD and 213 nm ultraviolet photodissociation (UVPD) facilitated identification of the chemical composition of the cross-links.
Combined EThcD and 213 nm UVPD facilitated identification of the chemical composition of cross-links in insulin dimers.
The combined utilization of EThcD and 213 nm ultraviolet photodissociation (UVPD) facilitated identification of the chemical composition of the cross-links.
Combined EThcD and 213 nm UVPD facilitated identification of the chemical composition of cross-links in insulin dimers.
The combined utilization of EThcD and 213 nm ultraviolet photodissociation (UVPD) facilitated identification of the chemical composition of the cross-links.
Combined EThcD and 213 nm UVPD facilitated identification of the chemical composition of cross-links in insulin dimers.
The combined utilization of EThcD and 213 nm ultraviolet photodissociation (UVPD) facilitated identification of the chemical composition of the cross-links.
Combined EThcD and 213 nm UVPD facilitated identification of the chemical composition of cross-links in insulin dimers.
The combined utilization of EThcD and 213 nm ultraviolet photodissociation (UVPD) facilitated identification of the chemical composition of the cross-links.
Combined EThcD and 213 nm UVPD facilitated identification of the chemical composition of cross-links in insulin dimers.
The combined utilization of EThcD and 213 nm ultraviolet photodissociation (UVPD) facilitated identification of the chemical composition of the cross-links.
Combined EThcD and 213 nm UVPD facilitated identification of the chemical composition of cross-links in insulin dimers.
The combined utilization of EThcD and 213 nm ultraviolet photodissociation (UVPD) facilitated identification of the chemical composition of the cross-links.
Combined EThcD and 213 nm UVPD facilitated identification of the chemical composition of cross-links in insulin dimers.
The combined utilization of EThcD and 213 nm ultraviolet photodissociation (UVPD) facilitated identification of the chemical composition of the cross-links.
Combined EThcD and 213 nm UVPD facilitated identification of the chemical composition of cross-links in insulin dimers.
The combined utilization of EThcD and 213 nm ultraviolet photodissociation (UVPD) facilitated identification of the chemical composition of the cross-links.
Combined EThcD and 213 nm UVPD facilitated identification of the chemical composition of cross-links in insulin dimers.
The combined utilization of EThcD and 213 nm ultraviolet photodissociation (UVPD) facilitated identification of the chemical composition of the cross-links.
Combined EThcD and 213 nm UVPD facilitated identification of the chemical composition of cross-links in insulin dimers.
The combined utilization of EThcD and 213 nm ultraviolet photodissociation (UVPD) facilitated identification of the chemical composition of the cross-links.
Combined EThcD and 213 nm UVPD facilitated identification of the chemical composition of cross-links in insulin dimers.
The combined utilization of EThcD and 213 nm ultraviolet photodissociation (UVPD) facilitated identification of the chemical composition of the cross-links.
Combined EThcD and 213 nm UVPD facilitated identification of the chemical composition of cross-links in insulin dimers.
The combined utilization of EThcD and 213 nm ultraviolet photodissociation (UVPD) facilitated identification of the chemical composition of the cross-links.
Residue-level identification of cross-link sites between chains was achievable for both dimers with MS3 analysis of MS2 fragments cleaved at the cross-link or additionally the interchain disulfide bonds.
Identification of cross-link sites between chains at residue level was achievable for both dimers with MS3 analysis of MS2 fragments cleaved at the cross-link or additionally the interchain disulfide bonds.
Residue-level identification of cross-link sites between chains was achievable for both dimers with MS3 analysis of MS2 fragments cleaved at the cross-link or additionally the interchain disulfide bonds.
Identification of cross-link sites between chains at residue level was achievable for both dimers with MS3 analysis of MS2 fragments cleaved at the cross-link or additionally the interchain disulfide bonds.
Residue-level identification of cross-link sites between chains was achievable for both dimers with MS3 analysis of MS2 fragments cleaved at the cross-link or additionally the interchain disulfide bonds.
Identification of cross-link sites between chains at residue level was achievable for both dimers with MS3 analysis of MS2 fragments cleaved at the cross-link or additionally the interchain disulfide bonds.
Residue-level identification of cross-link sites between chains was achievable for both dimers with MS3 analysis of MS2 fragments cleaved at the cross-link or additionally the interchain disulfide bonds.
Identification of cross-link sites between chains at residue level was achievable for both dimers with MS3 analysis of MS2 fragments cleaved at the cross-link or additionally the interchain disulfide bonds.
Residue-level identification of cross-link sites between chains was achievable for both dimers with MS3 analysis of MS2 fragments cleaved at the cross-link or additionally the interchain disulfide bonds.
Identification of cross-link sites between chains at residue level was achievable for both dimers with MS3 analysis of MS2 fragments cleaved at the cross-link or additionally the interchain disulfide bonds.
Residue-level identification of cross-link sites between chains was achievable for both dimers with MS3 analysis of MS2 fragments cleaved at the cross-link or additionally the interchain disulfide bonds.
Identification of cross-link sites between chains at residue level was achievable for both dimers with MS3 analysis of MS2 fragments cleaved at the cross-link or additionally the interchain disulfide bonds.
Residue-level identification of cross-link sites between chains was achievable for both dimers with MS3 analysis of MS2 fragments cleaved at the cross-link or additionally the interchain disulfide bonds.
Identification of cross-link sites between chains at residue level was achievable for both dimers with MS3 analysis of MS2 fragments cleaved at the cross-link or additionally the interchain disulfide bonds.
Residue-level identification of cross-link sites between chains was achievable for both dimers with MS3 analysis of MS2 fragments cleaved at the cross-link or additionally the interchain disulfide bonds.
Identification of cross-link sites between chains at residue level was achievable for both dimers with MS3 analysis of MS2 fragments cleaved at the cross-link or additionally the interchain disulfide bonds.
Residue-level identification of cross-link sites between chains was achievable for both dimers with MS3 analysis of MS2 fragments cleaved at the cross-link or additionally the interchain disulfide bonds.
Identification of cross-link sites between chains at residue level was achievable for both dimers with MS3 analysis of MS2 fragments cleaved at the cross-link or additionally the interchain disulfide bonds.
Residue-level identification of cross-link sites between chains was achievable for both dimers with MS3 analysis of MS2 fragments cleaved at the cross-link or additionally the interchain disulfide bonds.
Identification of cross-link sites between chains at residue level was achievable for both dimers with MS3 analysis of MS2 fragments cleaved at the cross-link or additionally the interchain disulfide bonds.
Top-down mass spectrometry workflows can identify cross-link types and sites in covalent insulin dimers induced by Fe2+ incubation or UV light stress.
we here demonstrate on two case studies of covalent insulin dimers, induced by Fe2+ incubation or ultraviolet (UV) light stress, that de novo characterization in top-down mass spectrometry (MS) workflows can identify cross-link types and sites
Top-down mass spectrometry workflows can identify cross-link types and sites in covalent insulin dimers induced by Fe2+ incubation or UV light stress.
we here demonstrate on two case studies of covalent insulin dimers, induced by Fe2+ incubation or ultraviolet (UV) light stress, that de novo characterization in top-down mass spectrometry (MS) workflows can identify cross-link types and sites
Top-down mass spectrometry workflows can identify cross-link types and sites in covalent insulin dimers induced by Fe2+ incubation or UV light stress.
we here demonstrate on two case studies of covalent insulin dimers, induced by Fe2+ incubation or ultraviolet (UV) light stress, that de novo characterization in top-down mass spectrometry (MS) workflows can identify cross-link types and sites
Top-down mass spectrometry workflows can identify cross-link types and sites in covalent insulin dimers induced by Fe2+ incubation or UV light stress.
we here demonstrate on two case studies of covalent insulin dimers, induced by Fe2+ incubation or ultraviolet (UV) light stress, that de novo characterization in top-down mass spectrometry (MS) workflows can identify cross-link types and sites
Top-down mass spectrometry workflows can identify cross-link types and sites in covalent insulin dimers induced by Fe2+ incubation or UV light stress.
we here demonstrate on two case studies of covalent insulin dimers, induced by Fe2+ incubation or ultraviolet (UV) light stress, that de novo characterization in top-down mass spectrometry (MS) workflows can identify cross-link types and sites
Top-down mass spectrometry workflows can identify cross-link types and sites in covalent insulin dimers induced by Fe2+ incubation or UV light stress.
we here demonstrate on two case studies of covalent insulin dimers, induced by Fe2+ incubation or ultraviolet (UV) light stress, that de novo characterization in top-down mass spectrometry (MS) workflows can identify cross-link types and sites
Top-down mass spectrometry workflows can identify cross-link types and sites in covalent insulin dimers induced by Fe2+ incubation or UV light stress.
we here demonstrate on two case studies of covalent insulin dimers, induced by Fe2+ incubation or ultraviolet (UV) light stress, that de novo characterization in top-down mass spectrometry (MS) workflows can identify cross-link types and sites
Top-down mass spectrometry workflows can identify cross-link types and sites in covalent insulin dimers induced by Fe2+ incubation or UV light stress.
we here demonstrate on two case studies of covalent insulin dimers, induced by Fe2+ incubation or ultraviolet (UV) light stress, that de novo characterization in top-down mass spectrometry (MS) workflows can identify cross-link types and sites
Top-down mass spectrometry workflows can identify cross-link types and sites in covalent insulin dimers induced by Fe2+ incubation or UV light stress.
we here demonstrate on two case studies of covalent insulin dimers, induced by Fe2+ incubation or ultraviolet (UV) light stress, that de novo characterization in top-down mass spectrometry (MS) workflows can identify cross-link types and sites
Top-down mass spectrometry workflows can identify cross-link types and sites in covalent insulin dimers induced by Fe2+ incubation or UV light stress.
we here demonstrate on two case studies of covalent insulin dimers, induced by Fe2+ incubation or ultraviolet (UV) light stress, that de novo characterization in top-down mass spectrometry (MS) workflows can identify cross-link types and sites
HCD, EThcD, and UVPD combined with MS3 were powerful tools for direct de novo characterization of cross-linked insulin dimers.
Our results demonstrated that higher-energy collisional dissociation (HCD), EThcD, and UVPD combined with MS3 were powerful tools for direct de novo characterization of cross-linked insulin dimers.
HCD, EThcD, and UVPD combined with MS3 were powerful tools for direct de novo characterization of cross-linked insulin dimers.
Our results demonstrated that higher-energy collisional dissociation (HCD), EThcD, and UVPD combined with MS3 were powerful tools for direct de novo characterization of cross-linked insulin dimers.
HCD, EThcD, and UVPD combined with MS3 were powerful tools for direct de novo characterization of cross-linked insulin dimers.
Our results demonstrated that higher-energy collisional dissociation (HCD), EThcD, and UVPD combined with MS3 were powerful tools for direct de novo characterization of cross-linked insulin dimers.
HCD, EThcD, and UVPD combined with MS3 were powerful tools for direct de novo characterization of cross-linked insulin dimers.
Our results demonstrated that higher-energy collisional dissociation (HCD), EThcD, and UVPD combined with MS3 were powerful tools for direct de novo characterization of cross-linked insulin dimers.
HCD, EThcD, and UVPD combined with MS3 were powerful tools for direct de novo characterization of cross-linked insulin dimers.
Our results demonstrated that higher-energy collisional dissociation (HCD), EThcD, and UVPD combined with MS3 were powerful tools for direct de novo characterization of cross-linked insulin dimers.
HCD, EThcD, and UVPD combined with MS3 were powerful tools for direct de novo characterization of cross-linked insulin dimers.
Our results demonstrated that higher-energy collisional dissociation (HCD), EThcD, and UVPD combined with MS3 were powerful tools for direct de novo characterization of cross-linked insulin dimers.
HCD, EThcD, and UVPD combined with MS3 were powerful tools for direct de novo characterization of cross-linked insulin dimers.
Our results demonstrated that higher-energy collisional dissociation (HCD), EThcD, and UVPD combined with MS3 were powerful tools for direct de novo characterization of cross-linked insulin dimers.
HCD, EThcD, and UVPD combined with MS3 were powerful tools for direct de novo characterization of cross-linked insulin dimers.
Our results demonstrated that higher-energy collisional dissociation (HCD), EThcD, and UVPD combined with MS3 were powerful tools for direct de novo characterization of cross-linked insulin dimers.
HCD, EThcD, and UVPD combined with MS3 were powerful tools for direct de novo characterization of cross-linked insulin dimers.
Our results demonstrated that higher-energy collisional dissociation (HCD), EThcD, and UVPD combined with MS3 were powerful tools for direct de novo characterization of cross-linked insulin dimers.
HCD, EThcD, and UVPD combined with MS3 were powerful tools for direct de novo characterization of cross-linked insulin dimers.
Our results demonstrated that higher-energy collisional dissociation (HCD), EThcD, and UVPD combined with MS3 were powerful tools for direct de novo characterization of cross-linked insulin dimers.
HCD, EThcD, and UVPD combined with MS3 were powerful tools for direct de novo characterization of cross-linked insulin dimers.
Our results demonstrated that higher-energy collisional dissociation (HCD), EThcD, and UVPD combined with MS3 were powerful tools for direct de novo characterization of cross-linked insulin dimers.
HCD, EThcD, and UVPD combined with MS3 were powerful tools for direct de novo characterization of cross-linked insulin dimers.
Our results demonstrated that higher-energy collisional dissociation (HCD), EThcD, and UVPD combined with MS3 were powerful tools for direct de novo characterization of cross-linked insulin dimers.
HCD, EThcD, and UVPD combined with MS3 were powerful tools for direct de novo characterization of cross-linked insulin dimers.
Our results demonstrated that higher-energy collisional dissociation (HCD), EThcD, and UVPD combined with MS3 were powerful tools for direct de novo characterization of cross-linked insulin dimers.
HCD, EThcD, and UVPD combined with MS3 were powerful tools for direct de novo characterization of cross-linked insulin dimers.
Our results demonstrated that higher-energy collisional dissociation (HCD), EThcD, and UVPD combined with MS3 were powerful tools for direct de novo characterization of cross-linked insulin dimers.
HCD, EThcD, and UVPD combined with MS3 were powerful tools for direct de novo characterization of cross-linked insulin dimers.
Our results demonstrated that higher-energy collisional dissociation (HCD), EThcD, and UVPD combined with MS3 were powerful tools for direct de novo characterization of cross-linked insulin dimers.
HCD, EThcD, and UVPD combined with MS3 were powerful tools for direct de novo characterization of cross-linked insulin dimers.
Our results demonstrated that higher-energy collisional dissociation (HCD), EThcD, and UVPD combined with MS3 were powerful tools for direct de novo characterization of cross-linked insulin dimers.
HCD, EThcD, and UVPD combined with MS3 were powerful tools for direct de novo characterization of cross-linked insulin dimers.
Our results demonstrated that higher-energy collisional dissociation (HCD), EThcD, and UVPD combined with MS3 were powerful tools for direct de novo characterization of cross-linked insulin dimers.
At the MS2 level, EThcD efficiently cleaved interchain disulfide bonds in insulin dimers to reveal cross-link connectivities between chains.
On the MS2 level, electron-transfer/higher-energy collision dissociation (EThcD) efficiently cleaved the interchain disulfide bonds in the dimers to reveal cross-link connectivities between chains.
At the MS2 level, EThcD efficiently cleaved interchain disulfide bonds in insulin dimers to reveal cross-link connectivities between chains.
On the MS2 level, electron-transfer/higher-energy collision dissociation (EThcD) efficiently cleaved the interchain disulfide bonds in the dimers to reveal cross-link connectivities between chains.
At the MS2 level, EThcD efficiently cleaved interchain disulfide bonds in insulin dimers to reveal cross-link connectivities between chains.
On the MS2 level, electron-transfer/higher-energy collision dissociation (EThcD) efficiently cleaved the interchain disulfide bonds in the dimers to reveal cross-link connectivities between chains.
At the MS2 level, EThcD efficiently cleaved interchain disulfide bonds in insulin dimers to reveal cross-link connectivities between chains.
On the MS2 level, electron-transfer/higher-energy collision dissociation (EThcD) efficiently cleaved the interchain disulfide bonds in the dimers to reveal cross-link connectivities between chains.
At the MS2 level, EThcD efficiently cleaved interchain disulfide bonds in insulin dimers to reveal cross-link connectivities between chains.
On the MS2 level, electron-transfer/higher-energy collision dissociation (EThcD) efficiently cleaved the interchain disulfide bonds in the dimers to reveal cross-link connectivities between chains.
At the MS2 level, EThcD efficiently cleaved interchain disulfide bonds in insulin dimers to reveal cross-link connectivities between chains.
On the MS2 level, electron-transfer/higher-energy collision dissociation (EThcD) efficiently cleaved the interchain disulfide bonds in the dimers to reveal cross-link connectivities between chains.
At the MS2 level, EThcD efficiently cleaved interchain disulfide bonds in insulin dimers to reveal cross-link connectivities between chains.
On the MS2 level, electron-transfer/higher-energy collision dissociation (EThcD) efficiently cleaved the interchain disulfide bonds in the dimers to reveal cross-link connectivities between chains.
At the MS2 level, EThcD efficiently cleaved interchain disulfide bonds in insulin dimers to reveal cross-link connectivities between chains.
On the MS2 level, electron-transfer/higher-energy collision dissociation (EThcD) efficiently cleaved the interchain disulfide bonds in the dimers to reveal cross-link connectivities between chains.
At the MS2 level, EThcD efficiently cleaved interchain disulfide bonds in insulin dimers to reveal cross-link connectivities between chains.
On the MS2 level, electron-transfer/higher-energy collision dissociation (EThcD) efficiently cleaved the interchain disulfide bonds in the dimers to reveal cross-link connectivities between chains.
At the MS2 level, EThcD efficiently cleaved interchain disulfide bonds in insulin dimers to reveal cross-link connectivities between chains.
On the MS2 level, electron-transfer/higher-energy collision dissociation (EThcD) efficiently cleaved the interchain disulfide bonds in the dimers to reveal cross-link connectivities between chains.
At the MS2 level, EThcD efficiently cleaved interchain disulfide bonds in insulin dimers to reveal cross-link connectivities between chains.
On the MS2 level, electron-transfer/higher-energy collision dissociation (EThcD) efficiently cleaved the interchain disulfide bonds in the dimers to reveal cross-link connectivities between chains.
At the MS2 level, EThcD efficiently cleaved interchain disulfide bonds in insulin dimers to reveal cross-link connectivities between chains.
On the MS2 level, electron-transfer/higher-energy collision dissociation (EThcD) efficiently cleaved the interchain disulfide bonds in the dimers to reveal cross-link connectivities between chains.
At the MS2 level, EThcD efficiently cleaved interchain disulfide bonds in insulin dimers to reveal cross-link connectivities between chains.
On the MS2 level, electron-transfer/higher-energy collision dissociation (EThcD) efficiently cleaved the interchain disulfide bonds in the dimers to reveal cross-link connectivities between chains.
At the MS2 level, EThcD efficiently cleaved interchain disulfide bonds in insulin dimers to reveal cross-link connectivities between chains.
On the MS2 level, electron-transfer/higher-energy collision dissociation (EThcD) efficiently cleaved the interchain disulfide bonds in the dimers to reveal cross-link connectivities between chains.
At the MS2 level, EThcD efficiently cleaved interchain disulfide bonds in insulin dimers to reveal cross-link connectivities between chains.
On the MS2 level, electron-transfer/higher-energy collision dissociation (EThcD) efficiently cleaved the interchain disulfide bonds in the dimers to reveal cross-link connectivities between chains.
At the MS2 level, EThcD efficiently cleaved interchain disulfide bonds in insulin dimers to reveal cross-link connectivities between chains.
On the MS2 level, electron-transfer/higher-energy collision dissociation (EThcD) efficiently cleaved the interchain disulfide bonds in the dimers to reveal cross-link connectivities between chains.
At the MS2 level, EThcD efficiently cleaved interchain disulfide bonds in insulin dimers to reveal cross-link connectivities between chains.
On the MS2 level, electron-transfer/higher-energy collision dissociation (EThcD) efficiently cleaved the interchain disulfide bonds in the dimers to reveal cross-link connectivities between chains.
In the Fe2+-induced insulin dimer, Phe1 from both B-chains were cross-linked through a -CH2- linkage.
In the Fe2+-induced dimer, Phe1 from both B-chains were cross-linked through a -CH2-.
In the Fe2+-induced insulin dimer, Phe1 from both B-chains were cross-linked through a -CH2- linkage.
In the Fe2+-induced dimer, Phe1 from both B-chains were cross-linked through a -CH2-.
In the Fe2+-induced insulin dimer, Phe1 from both B-chains were cross-linked through a -CH2- linkage.
In the Fe2+-induced dimer, Phe1 from both B-chains were cross-linked through a -CH2-.
In the Fe2+-induced insulin dimer, Phe1 from both B-chains were cross-linked through a -CH2- linkage.
In the Fe2+-induced dimer, Phe1 from both B-chains were cross-linked through a -CH2-.
In the Fe2+-induced insulin dimer, Phe1 from both B-chains were cross-linked through a -CH2- linkage.
In the Fe2+-induced dimer, Phe1 from both B-chains were cross-linked through a -CH2-.
In the Fe2+-induced insulin dimer, Phe1 from both B-chains were cross-linked through a -CH2- linkage.
In the Fe2+-induced dimer, Phe1 from both B-chains were cross-linked through a -CH2-.
In the Fe2+-induced insulin dimer, Phe1 from both B-chains were cross-linked through a -CH2- linkage.
In the Fe2+-induced dimer, Phe1 from both B-chains were cross-linked through a -CH2-.
In the Fe2+-induced insulin dimer, Phe1 from both B-chains were cross-linked through a -CH2- linkage.
In the Fe2+-induced dimer, Phe1 from both B-chains were cross-linked through a -CH2-.
In the Fe2+-induced insulin dimer, Phe1 from both B-chains were cross-linked through a -CH2- linkage.
In the Fe2+-induced dimer, Phe1 from both B-chains were cross-linked through a -CH2-.
In the Fe2+-induced insulin dimer, Phe1 from both B-chains were cross-linked through a -CH2- linkage.
In the Fe2+-induced dimer, Phe1 from both B-chains were cross-linked through a -CH2-.
In the UV light-induced insulin dimer, Tyr14 of the A-chain participated in an -O-S- cross-link in which the sulfur was derived either from Cys7 or Cys19 of the B-chain.
in the UV light-induced dimer, Tyr14 of the A-chain participated in an -O-S- cross-link in which the sulfur was derived either from Cys7 or Cys19 of the B-chain
In the UV light-induced insulin dimer, Tyr14 of the A-chain participated in an -O-S- cross-link in which the sulfur was derived either from Cys7 or Cys19 of the B-chain.
in the UV light-induced dimer, Tyr14 of the A-chain participated in an -O-S- cross-link in which the sulfur was derived either from Cys7 or Cys19 of the B-chain
In the UV light-induced insulin dimer, Tyr14 of the A-chain participated in an -O-S- cross-link in which the sulfur was derived either from Cys7 or Cys19 of the B-chain.
in the UV light-induced dimer, Tyr14 of the A-chain participated in an -O-S- cross-link in which the sulfur was derived either from Cys7 or Cys19 of the B-chain
In the UV light-induced insulin dimer, Tyr14 of the A-chain participated in an -O-S- cross-link in which the sulfur was derived either from Cys7 or Cys19 of the B-chain.
in the UV light-induced dimer, Tyr14 of the A-chain participated in an -O-S- cross-link in which the sulfur was derived either from Cys7 or Cys19 of the B-chain
In the UV light-induced insulin dimer, Tyr14 of the A-chain participated in an -O-S- cross-link in which the sulfur was derived either from Cys7 or Cys19 of the B-chain.
in the UV light-induced dimer, Tyr14 of the A-chain participated in an -O-S- cross-link in which the sulfur was derived either from Cys7 or Cys19 of the B-chain
In the UV light-induced insulin dimer, Tyr14 of the A-chain participated in an -O-S- cross-link in which the sulfur was derived either from Cys7 or Cys19 of the B-chain.
in the UV light-induced dimer, Tyr14 of the A-chain participated in an -O-S- cross-link in which the sulfur was derived either from Cys7 or Cys19 of the B-chain
In the UV light-induced insulin dimer, Tyr14 of the A-chain participated in an -O-S- cross-link in which the sulfur was derived either from Cys7 or Cys19 of the B-chain.
in the UV light-induced dimer, Tyr14 of the A-chain participated in an -O-S- cross-link in which the sulfur was derived either from Cys7 or Cys19 of the B-chain
In the UV light-induced insulin dimer, Tyr14 of the A-chain participated in an -O-S- cross-link in which the sulfur was derived either from Cys7 or Cys19 of the B-chain.
in the UV light-induced dimer, Tyr14 of the A-chain participated in an -O-S- cross-link in which the sulfur was derived either from Cys7 or Cys19 of the B-chain
In the UV light-induced insulin dimer, Tyr14 of the A-chain participated in an -O-S- cross-link in which the sulfur was derived either from Cys7 or Cys19 of the B-chain.
in the UV light-induced dimer, Tyr14 of the A-chain participated in an -O-S- cross-link in which the sulfur was derived either from Cys7 or Cys19 of the B-chain
In the UV light-induced insulin dimer, Tyr14 of the A-chain participated in an -O-S- cross-link in which the sulfur was derived either from Cys7 or Cys19 of the B-chain.
in the UV light-induced dimer, Tyr14 of the A-chain participated in an -O-S- cross-link in which the sulfur was derived either from Cys7 or Cys19 of the B-chain
Approval Evidence
1 source3 linked approval claimsfirst-pass slug electron-transfer-higher-energy-collision-dissociation
electron-transfer/higher-energy collision dissociation (EThcD)
Source:
method capabilitysupports
Combined EThcD and 213 nm UVPD facilitated identification of the chemical composition of cross-links in insulin dimers.
The combined utilization of EThcD and 213 nm ultraviolet photodissociation (UVPD) facilitated identification of the chemical composition of the cross-links.
Source:
method conclusionsupports
HCD, EThcD, and UVPD combined with MS3 were powerful tools for direct de novo characterization of cross-linked insulin dimers.
Our results demonstrated that higher-energy collisional dissociation (HCD), EThcD, and UVPD combined with MS3 were powerful tools for direct de novo characterization of cross-linked insulin dimers.
Source:
method observationsupports
At the MS2 level, EThcD efficiently cleaved interchain disulfide bonds in insulin dimers to reveal cross-link connectivities between chains.
On the MS2 level, electron-transfer/higher-energy collision dissociation (EThcD) efficiently cleaved the interchain disulfide bonds in the dimers to reveal cross-link connectivities between chains.
Source:
Comparisons
Source-backed strengths
The reported workflow could identify cross-link types and sites in covalent insulin dimers generated by two distinct stress conditions, indicating applicability across more than one dimer chemistry. Combined EThcD and 213 nm UVPD facilitated cross-link composition assignment, and MS3 of MS2 fragments cleaved at the cross-link or interchain disulfide bonds enabled residue-level interchain site identification for both dimers.
Source:
UVPD provided identification of cross-link sites in the Fe2+-induced dimer without MS3, while cross-link site identification with MS2 was not possible for the UV light-induced dimer.
Compared with electron-electron double resonance spectroscopy
electron-transfer/higher-energy collision dissociation and electron-electron double resonance spectroscopy address a similar problem space.
Shared frame: same top-level item type; shared mechanisms: heterodimerization; same primary input modality: light
Relative tradeoffs: looks easier to implement in practice.
Compared with top-down mass spectrometry
electron-transfer/higher-energy collision dissociation and top-down mass spectrometry address a similar problem space.
Shared frame: same top-level item type; shared mechanisms: heterodimerization, photocleavage; same primary input modality: light
Relative tradeoffs: looks easier to implement in practice.
Compared with ultraviolet photodissociation
electron-transfer/higher-energy collision dissociation and ultraviolet photodissociation address a similar problem space.
Shared frame: same top-level item type; shared mechanisms: heterodimerization, ultraviolet photodissociation; same primary input modality: light
Relative tradeoffs: looks easier to implement in practice.
Ranked Citations
- 1.